Cell nuclei were sonicated to shear DNA in 300 μl of sonication buffer, using a Bioruptor Plus (Diagenode) (18 times, 15 sec on /20 sec off each time, 9 W potency), obtaining lengths between 300 and 600 bp. Immunoprecipitations were carried out overnight at 4 °C using 40 μg of chromatin and 5 μg of SALL2 (anti-SALL2; Bethyl). The SALL2 total knockout was used as a negative control of antibody binding To prepare each library was used the KAPA Hiper Prep kit and 7 adapters from the KAPA SI Adapters kit, both according to the manufacturer's protocols. Briefly, the first step in library preparation was to convert any overhangs in the ChIP DNA into phosphorylated blunt ends. The 3′ ends were then adenylated, and adaptors ligated onto the ends of the fragments. The library size (348 bp) and the DNA concentration (14,486 nM) was obtained by using an Agilent 2100 Bioanalyzer